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Determination of apolipoprotein E genotypes by single-strand conformational polymorphism

M Y Tsai1, P Suess, K Schwichtenberg

  • 1Department of Laboratory Medicine and Pathology, University of Minnesota Medical School, Minneapolis 55455.

Clinical Chemistry
|October 1, 1993
PubMed
Summary

Single-strand conformational polymorphism (SSCP) effectively determines apolipoprotein E (Apo E) genotypes, including rare alleles. This method accurately identifies Apo E gene variations, aligning with other genotyping techniques.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Apolipoprotein E (Apo E) plays a crucial role in lipid metabolism and is associated with various diseases.
  • Accurate genotyping of Apo E alleles is essential for genetic studies and clinical applications.
  • Existing genotyping methods may have limitations in detecting rare Apo E alleles.

Purpose of the Study:

  • To evaluate the efficacy of single-strand conformational polymorphism (SSCP) for determining apolipoprotein E (Apo E) genotypes.
  • To assess the utility of SSCP in identifying both common and rare Apo E alleles.
  • To compare SSCP results with established genotyping methods.

Main Methods:

  • Single-strand conformational polymorphism (SSCP) analysis was performed on a 295-base-pair (bp) DNA fragment encoding amino acid residues 80-178 of the Apo E protein.

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  • Genotyping was conducted on 47 individuals.
  • Comparison with PCR-based amplification refractory mutation system (ARMS) genotyping was performed for five individuals with discrepant phenotyping results.

    • Main Results:

    • SSCP analysis generated distinct patterns for the three common Apo E alleles.
    • SSCP results for Apo E genotyping were consistent with ARMS genotyping, even in cases with differing phenotyping results.
    • The SSCP method demonstrated effectiveness in identifying Apo E alleles, including those involving mutations in positions 120-160, which are associated with rare alleles.

    Conclusions:

    • Single-strand conformational polymorphism (SSCP) is a reliable and effective method for determining apolipoprotein E (Apo E) genotypes.
    • The SSCP approach is valuable for identifying both common and rare Apo E alleles, offering an advantage over methods that may miss rare variants.
    • This SSCP-based genotyping strategy provides a robust tool for Apo E genetic analysis in research and potentially clinical settings.